Update on sputum methodology.

Over the past few years cellular and biochemical sputum examinations have become important instruments to assess airway inflammation. The aim of this review is to summarize new methodological developments and aspects, which are currently under investigation. The use of isotonic saline has increased safety of inductions in patients with severe asthma and in children. The origin of sputum is better understood, as is the need to standardize the volume and duration of induction. It also needs to be borne in mind that the induction procedure itself is able to cause changes in sputum composition. However, the basic induction and processing procedures have not changed much over the last few years, and therefore the method is still time consuming. The analysis of ECP in lysed sputum cells as a marker for the number of eosinophils has been suggested to overcome this problem, but needs further validation. Furthermore, storage of sputum has been studied, as well as early fixation or freezing of sputum cells to elongate the time between induction and processing. Differential cell counts by flow cytometry are still difficult, but the method has increased knowledge concerning lymphocyte subsets and the activation status of sputum cells. The use of induced sputum to noninvasively measure airway inflammation in clinical trials will offer additional information, but the proper use and interpretation of sputum outcome parameters will need further investigation.